T84 monolayers are superior to Caco-2 as a model system of colonocytes
T84 monolayers are superior to Caco-2 as a model system of colonocytes
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DOI：
10.1007/s00418-017-1539-7
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发表时间：
2017-07-01
影响因子：
2.3
通讯作者：
Laukens, Debby
中科院分区：
文献类型：
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作者：
Devriese, Sarah;Van den Bossche, Lien;Laukens, Debby
结肠腺癌衍生的Caco-2和T84上皮细胞系经常用作功能性上皮屏障的体外模型系统。两者可互换使用，尽管有证据表明分化的Caco-2细胞比结肠细胞更容易让人联想到小肠肠上皮细胞，而分化的T84细胞则不太清楚。因此，本研究的目的是进一步鉴定和比较分化的Caco-2和T84细胞。目的是（1）比较刷状缘形态，（2）测量肠细胞和结肠细胞特异性基因的表达，（3）比较它们对丁酸盐的反应，丁酸盐依赖于单羧酸转运蛋白1（MCT 1），一种主要在结肠细胞中表达的顶端蛋白。T84微绒毛明显短于Caco-2细胞，这是小肠上皮细胞与结肠上皮细胞的特征性差异。此外，在分化的Caco-2细胞中表达的肠上皮细胞相关刷状缘酶在T84成熟过程中没有增加，而结肠标记物如MCT 1在分化的T84细胞中比分化的Caco-2细胞更丰富。因此，T84细胞显示出对丁酸盐的屏障功能的剂量响应性改善，这在Caco-2细胞中不存在。另一方面，Caco-2和T84单层之间上皮Toll样受体表达的差异并没有导致相应的差异功能反应。我们的结论是，分化的Caco-2和T84细胞具有不同的形态，生化和功能特性，这表明T84细胞不像Caco-2细胞那样获得成熟小肠肠上皮细胞的生化特征，但在整个分化过程中保留了许多原始结肠特征。这些发现可以帮助研究人员选择合适的肠上皮细胞系用于特定的体外研究目的。
Colonic adenocarcinoma-derived Caco-2 and T84 epithelial cell lines are frequently used as in vitro model systems of functional epithelial barriers. Both are utilised interchangeably despite evidence that differentiated Caco-2 cells are more reminiscent of small intestinal enterocytes than of colonocytes, whereas differentiated T84 cells are less well characterised. The aim of this study was, therefore, to further characterise and compare differentiated Caco-2 and T84 cells. The objectives were to (1) compare the brush border morphology, (2) measure the expression of enterocyte- and colonocyte-specific genes and (3) compare their response to butyrate, which is dependent on the monocarboxylate transporter 1 (MCT1), an apical protein expressed primarily in colonocytes. T84 microvilli were significantly shorter than those of Caco-2 cells, which is a characteristic difference between small intestinal enterocytes and colonocytes. Also, enterocyte-associated brush border enzymes expressed in differentiated Caco-2 cells were not increased during T84 maturation, whereas colonic markers such as MCT1 were more abundant in differentiated T84 cells compared to differentiated Caco-2 cells. Consequently, T84 cells displayed a dose-responsive improvement of barrier function towards butyrate, which was absent in Caco-2 cells. On the other hand, differences in epithelial toll-like receptor expression between Caco-2 and T84 monolayers did not result in a corresponding differential functional response. We conclude that differentiated Caco-2 and T84 cells have distinct morphological, biochemical and functional characteristics, suggesting that T84 cells do not acquire the biochemical signature of mature small intestinal enterocytes like Caco-2 cells, but retain much of their original colonic characteristics throughout differentiation. These findings can help investigators select the appropriate intestinal epithelial cell line for specific in vitro research purposes.